GSM7049559: Kidney, epithelial cell, KOhighF2; Mus musculus; RNA-Seq
According to protocol using trizol from invitrogen The RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina  using manufacturer's instructions (New England Biolabs, Ipswich, MA, USA). Briefly, mRNAs were initially  enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand  and second strand cDNA were subsequently synthesized. cDNA fragments were end repaired and  adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition  and library enrichment by PCR with limited cycles. The sequencing libraries were validated on the Agilent  TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 2.0 Fluorometer  (ThermoFisher Scientific, Waltham, MA, USA) as well as by quantitative PCR (KAPA Biosystems,  Wilmington, MA, USA).  The sequencing libraries were multiplexed and clustered onto a flowcell. After clustering, the flowcell was  loaded onto the Illumina HiSeq instrument according to manufacturer's instructions. The samples were  sequenced using a 2x150bp Paired End (PE) configuration. Image analysis and base calling were  conducted by the HiSeq Control Software (HCS). Raw sequence data (.bcl files) generated from Illumina  HiSeq was converted into fastq files and de-multiplexed using Illumina bcl2fastq 2.20 software. One mis-  match was allowed for index sequence identification.